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In vitro mutagenesis of the GGS1/TPS1 gene of Sacharomyces cerevisiae : graduation thesis = In vitro mutacija gena GGS1/TPS1 kvasovke Saccharomyces cerevisiae : diplomska naloga
Buzeti, Tatjana
In vitro mutacije na genu GGS1/TPS1 kvasovke Saccharomyces cerevisiae so potekale z naključno in s ciljano mutagenezo. Rezultat ciljane mutageneze so bile 3 mutacije, D395A, H185A in N295A. S ...plazmidi Ycplac33/TPS1, ki so vsebovali mutagene alele, sem transformirala seve W303-1Atps1[delta] in W303-1Atps1[delta]tps2[delta]. Na podlagi opazovanja fenotipskih sprememb mutantov na selektivnem gojišču sem opazovala spremembo v glikolitičnem fluksu oz. njihovo rast na glukozi ter termotoleranco. Aktivnost trehaloza-6-fosfat sintaze (aktivnost Tps1) in vsebnost trehaloze sem spremljala z biokemičnimi metodami. Pri vseh treh mutacijah je bila popolnoma uničena aktivnost Tps1, posledica česar je tudi minimalna vsebnost trehaloze v primeru mutacij D395A in H185A. V primeru mutacije N295A je bila vsebnost trehaloze nekoliko višja, s čimer bi lahko razlagali še vedno obstoječo regulacijo glukoznega fluksa v glikolizo in s tem rast na glukoznem gojišču. Rezultati potrjujejo vlogo gena GGS1/TPS1 v glikolitičnem fluksu, vendar ne moremo potrditi niti zavreči nobene izmed treh predlaganih razlag njegove regulatorne vloge v omenjeni metabolni poti.
Type of material - undergraduate thesis
; adult, serious
Publication and manufacture - Ljubljana : [BF, Oddelek za živilstvo] : [T. Buzeti], 1999
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